Analyzing AntibodyFc-Receptor Interactions

Cellular receptors for immunoglobulins (Fc-receptors; FcR) are central mediators of antibody-triggered effector functions. Immune complex (IC) binding to FcRs results in a variety of reactions such as the release of inflammatory mediators, antibody dependent cellular cytotoxicity (ADCC) and phagocytosis of ICs. Analyzing antibody–FcR (Ab–FcR) interactions in vitro is essential to determine the effector mechanisms, binding characteristics and affinity parameters that will impact and predict antibody activity in vivo. The methods described in this chapter include the generation of ICs and soluble FcR variants, as well as ELISA and FACS-based assays to study Ab–FcR interactions.